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LRRN1 / Leucine-rich repeat neuronal protein 1 · IHC design guide
Design Immunohistochemistry for LRRN1
Plan LRRN1 chromogenic IHC in paraffin sections using high-staining fallopian tube glandular cells or skeletal muscle myocytes as positive tissue references (HPA tissue IHC). Compare cytoplasmic staining with cells reported as undetected, such as adipocytes, while accounting for medium consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LRRN1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | General cytoplasmic staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in positive cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A12074-1) | |
| Positive control | Fallopian tube+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA have medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | 0 isoforms annotated; epitope may be extracellular or cytoplasmic (UniProt) |
Section 1
Recommended LRRN1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (A12074-1 datasheet). Two published LRRN1 IHC studies provide sample preparation or detection details (PMC8479295; PMC6609808).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A12074-1) |
| Fixation | Image fixative and duration unreported (datasheet A12074-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A12074-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A12074-1) |
| Primary antibody | Rabbit anti-LRRN1, 2-5 μg/ml (datasheet A12074-1) |
| Primary incubation | Overnight at 4 °C (datasheet A12074-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A12074-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LRRN1-positive staining in glandular cells of fallopian tube (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (A12074-1 datasheet); neither article excerpt specifies retrieval conditions (PMC8479295; PMC6609808).
Section 2
What Is the Expected LRRN1 Staining Pattern?
In paraffin-section IHC, expect general cytoplasmic LRRN1 staining, with high staining reported in fallopian tube glandular cells, skeletal-muscle myocytes, testis spermatogonia and tonsil squamous epithelial cells (HPA: tissue IHC). LRRN1 is a membrane protein with an extracellular region at residues 26–631 and a transmembrane segment at 632–652 (UniProt Q6UXK5 topology). HPA rates the tissue IHC evidence Enhanced, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Cytoplasmic staining is strongest in the expected cells, while nearby cells vary in intensity. | This fits HPA’s general cytoplasmic IHC profile. High staining is reported in fallopian tube glandular cells, myocytes, spermatogonia and tonsil squamous epithelial cells; medium staining is reported in caudate glial and cerebellar Purkinje cells (HPA: tissue IHC). Compare the named cell population, not just the tissue label, when judging a section. |
| Predominantly nuclear staining appears in cells expected to stain. | A dominant nuclear pattern conflicts with HPA’s general cytoplasmic IHC profile and UniProt’s membrane annotation (HPA: tissue IHC; UniProt Q6UXK5 subcellular location). Treat it as suspect until a positive-control section and appropriate detection controls support specificity. The available sources do not establish a nuclear IHC pattern for LRRN1. |
| Strong staining appears in cells reported as not detected, such as adipocytes or lymph-node germinal-center cells. | That distribution conflicts with the named HPA observations (HPA: not detected in adipocytes and lymph-node germinal-center cells). Cross-reactivity or endogenous detection activity are possible explanations, not diagnoses from appearance alone (general IHC practice). Check a matched positive section and a primary-antibody-omission control before assigning the signal to LRRN1. |
| Diffuse color covers tissue, cell-free spaces or much of the section without a clear cell pattern. | This does not reproduce the cell-specific HPA observations or its general cytoplasmic profile (HPA: tissue IHC). Background from detection chemistry or insufficient blocking or washing can obscure interpretation (general IHC practice). Judge the signal only after a control section shows that the diffuse color is acceptably low. |
| No staining appears in fallopian tube glandular cells or testis spermatogonia. | Both are reported as high-staining populations, so an entirely blank result warrants a technical check (HPA: High in fallopian tube glandular cells and testis spermatogonia). HPA’s Enhanced rating does not guarantee signal in every preparation (HPA: medium staining–RNA consistency). Verify tissue identity, detection reagents and the validated IHC conditions before calling the specimen negative. |
💡Expected LRRN1 appearanceCall a section positive when the expected cell population shows discernible, generally cytoplasmic chromogenic staining—potentially high in fallopian tube glandular cells, myocytes, spermatogonia or tonsil squamous epithelium—while dominant nuclear color or strong staining in HPA not-detected cell populations calls for control review (HPA: tissue IHC; UniProt Q6UXK5 subcellular location).
| Cell population and comparator tissue | HPA scores specific cells: tonsil squamous epithelial cells are High, whereas oral-mucosa squamous epithelial cells are Not detected (HPA: tissue IHC). Do not transfer an intensity expectation between tissues solely because their cell labels match. Use the annotated cell population as the comparison point. |
| Protein topology and epitope location | The mature chain begins at residue 26; UniProt places residues 26–631 extracellularly and 653–716 in the cytoplasmic region (UniProt Q6UXK5 topology and processing). Antibody epitope location is not supplied, so topology cannot predict this antibody’s staining intensity or prescribe retrieval conditions. |
| Antibody validation and staining confidence | HPA lists rabbit polyclonal HPA011071 as IHC Enhanced; its tissue page describes medium consistency between antibody staining and RNA expression (HPA: antibodies; HPA: tissue IHC reliability). This supports using its observed pattern as a reference while retaining section-level controls for discordant results. |
| IF/ICC question: where is LRRN1 seen? | HPA reports mainly endoplasmic-reticulum localization, with additional Golgi, vesicle, centriolar-satellite and basal-body localization in ICC-IF (HPA: subcellular ICC-IF). That is a separate cell-imaging observation; interpret paraffin-section chromogenic staining against HPA’s general cytoplasmic tissue IHC profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported High population is blank in the test section. | A failed staining run or a section lacking the annotated cell population can explain the discrepancy; HPA reports High staining in fallopian tube glandular cells and testis spermatogonia (HPA: tissue IHC). | Confirm the cell population on the counterstained section, then check an HPA-reported positive tissue stained in the same run. Review the antibody’s IHC-validated conditions and detection steps before interpreting absence as biology (general IHC practice). |
| All sections, including the primary-antibody-omission control, show chromogenic color. | Signal present without primary antibody points to background from the detection workflow, including possible endogenous enzyme activity; it cannot establish LRRN1 staining (general IHC practice). | Check the detection system’s blocking and control steps, then repeat the matched control. Score LRRN1 only when the primary-antibody section has a distinct cell-associated pattern above control background (general IHC practice). |
| Color is widespread, but expected cells cannot be distinguished. | Diffuse background can mask the cell-specific comparison needed for HPA’s tissue IHC pattern (HPA: tissue IHC; general IHC practice). | Inspect the omission control and review blocking, washing and chromogen development. Compare expected positive cells with adjacent cells only after background permits a clear distinction (general IHC practice). |
| A section shows mainly nuclear staining. | That compartment differs from HPA’s general cytoplasmic tissue IHC profile and UniProt’s membrane annotation (HPA: tissue IHC; UniProt Q6UXK5 subcellular location). | Review morphology and the omission control, then compare with a same-run positive section. If the nuclear pattern persists without the expected cytoplasmic distribution, report it as discordant rather than confirmed LRRN1 localization (general IHC practice). |
| Strong signal appears in an HPA not-detected cell population. | Cross-reactivity or endogenous detection activity may account for unexpected color; HPA reports adipocytes and lymph-node germinal-center cells as Not detected (HPA: tissue IHC; general IHC practice). | Identify the stained cells on the counterstain, run a primary-antibody-omission control and compare an HPA-reported positive population in the same staining run. Avoid treating the unexpected signal alone as evidence of LRRN1 (general IHC practice). |
Experiment evidence
IHC & IF images showing LRRN1 staining
A12074-1 · IHCIHC analysis of LRRN1 using anti-LRRN1 antibody (A12074-1). LRRN1 was detected in a paraffin-embedded section of rat brain tissue. Heat medi
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000175928-LRRN1/tissue/cerebral+cortex#imid_3308808
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000175928-LRRN1/tissue/colon#imid_3308734
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000175928-LRRN1/tissue/liver#imid_3308735
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000175928-LRRN1/tissue/kidney#imid_3308736
HPA · ICC-IF AF22Human Protein Atlas immunofluorescence, AF22, antibody HPA011071; green = LRRN1, blue = nucleus, red = microtubules.
HPA · ICC-IF AF22Human Protein Atlas immunofluorescence, AF22, antibody HPA011071; green = LRRN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LRRN1 IHC & IF
🧪Run fallopian tube first and require staining in glandular cells (HPA: High in fallopian tube glandular cells). Use prostate glandular cells as a tissue negative (HPA: Not detected in prostate glandular cells); on the positive slide, treat cells outside the glandular compartment as internal negative comparators only if they remain unstained.
Positive control tissue: Fallopian tube (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LRRN1 in AF22, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls for the rabbit catalog antibody (selected-SKU IHC caption: rabbit primary). Confirm target-dependent staining with LRRN1 knockout material or a validated immunizing-peptide block, if available. Block endogenous peroxidase and assess nonspecific DAB signal in the fallopian tube section (standard chromogenic IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected-SKU paraffin-section caption does not state the fixative (selected-SKU IHC caption: fixative unreported). Heat retrieval with EDTA at pH 8.0 was used for rat brain paraffin IHC, so its need and conditions in fallopian tube require optimization (selected-SKU IHC caption: rat brain, EDTA retrieval). The evidence does not establish that frozen sections or IF are easier; in fallopian tube glands, score cell-associated staining separately from luminal DAB deposits or background (standard IHC interpretation practice).
HPA tissue IHC evidence for LRRN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Tonsil | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LRRN1 IHC Tips
Troubleshoot LRRN1 staining in paraffin sections by checking retrieval, tissue morphology, controls, and the cell type and compartment carrying the signal.
Which retrieval condition should I start with for LRRN1 IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A12074-1). The rat brain image for this antibody used that buffer, followed by 2 μg/mL primary antibody overnight at 4°C (A12074-1 tissue-IHC caption). Keep heating and cooling conditions identical across slides, and include a no-primary control to identify retrieval-associated background (standard IHC practice). If staining is weak, compare shorter and longer heat exposure on adjacent sections before trying another buffer, since excessive heating can damage morphology (standard IHC practice). Assess signal alongside intact cell morphology rather than accepting diffuse DAB alone as specific staining (standard IHC practice).
Could fixation explain variable LRRN1 staining between paraffin sections?
The selected rat brain specimen is described as paraffin embedded, but its fixative and fixation duration are unreported (A12074-1 tissue-IHC caption). LRRN1-specific sensitivity to fixative type, concentration, delay, or duration is therefore unknown (A12074-1 tissue-IHC caption). For prospective specimens, record those variables and keep processing consistent across comparison groups (standard IHC practice). When staining differs, process serial sections together using EDTA at pH 8.0, the same antibody concentration, and identical detection conditions (datasheet A12074-1; standard IHC practice). Check morphology and a no-primary control before attributing a DAB difference to antigen preservation (standard IHC practice).
Should LRRN1 appear at the cell surface or in the cytoplasm?
LRRN1 is a membrane protein with an extracellular region at residues 26–631, a transmembrane segment at 632–652, and a cytoplasmic tail at 653–716 (UniProt Q6UXK5 topology). Tissue IHC is described as generally cytoplasmic, while subcellular imaging places it mainly in the endoplasmic reticulum, with additional Golgi and vesicle localisation (HPA tissue IHC; HPA subcellular). Accordingly, cytoplasmic DAB does not by itself contradict membrane topology (UniProt Q6UXK5 topology; HPA tissue IHC). Compare staining with cellular outlines and organelle-rich cytoplasm, using adjacent sections and consistent counterstaining to resolve boundaries (standard IHC practice). Avoid assigning a specific organelle from chromogenic staining alone (standard IHC practice).
How does an unknown antibody epitope affect LRRN1 IHC troubleshooting?
The supplied record annotates 0 isoforms but does not identify the catalog antibody’s epitope (UniProt Q6UXK5 record; A12074-1 tissue-IHC caption). LRRN1 has an extracellular region at residues 26–631 and a cytoplasmic tail at 653–716, so epitope location would affect how staining is interpreted (UniProt Q6UXK5 topology). Its annotated glycosylation sites are 96, 117, 385, and 517; their effect on this antibody’s staining has not been established by the supplied evidence (UniProt Q6UXK5 glycosylation; A12074-1 tissue-IHC caption). If staining is unexpected, request epitope information before assigning a domain-specific mechanism (standard IHC practice). Compare retrieval conditions using matched sections and appropriate controls (standard IHC practice).
How should I investigate LRRN1 localisation by multiplex immunofluorescence?
Pair LRRN1 with a validated marker for the cell population being examined, such as Purkinje cells in cerebellum, which show medium tissue-IHC staining (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel when tissue autofluorescence obscures shorter wavelengths (standard IF practice). The antibody epitope is unspecified, while LRRN1 has extracellular residues 26–631 and a cytoplasmic tail at 653–716 (A12074-1 tissue-IHC caption; UniProt Q6UXK5 topology). Compare no permeabilisation with mild permeabilisation: access to a cytoplasmic epitope generally requires membrane permeabilisation (standard IF practice). Include single-label and no-primary controls when judging colocalisation (standard IF practice).
What should I check when LRRN1 DAB staining is widespread?
First compare a no-primary slide with the stained section to identify signal arising from detection chemistry or tissue rather than the primary antibody (standard IHC practice). Block endogenous peroxidase before HRP detection and inspect pigment or precipitate under the microscope (standard IHC practice). The selected rat brain workflow used 10% goat serum before 2 μg/mL primary antibody and an HRP/DAB readout (A12074-1 tissue-IHC caption). If background persists, compare a lower primary concentration and longer washes while keeping retrieval at EDTA pH 8.0 (datasheet A12074-1; standard IHC practice). Exclude folds, damaged edges, and necrotic regions from interpretation (standard IHC practice).
How can I quantify LRRN1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and cell population before scoring, because reported staining varies by cell type and tissue (HPA tissue IHC; standard IHC practice). For a cell-based endpoint, report the percentage of positive cells and an H-score: sum each intensity grade from 0–3 multiplied by its percentage of cells, giving a 0–300 scale (standard IHC practice). Count morphologically identified cells and normalise positive counts to all eligible cells in the same compartment (standard IHC practice). If measuring stained area, report positive area per mm² of viable tissue (standard IHC practice). Keep section thickness, imaging, DAB development, and scoring thresholds consistent across groups (standard IHC practice).
How can I distinguish true LRRN1 signal from a staining artefact?
Look for reproducible staining in morphologically intact cells, with a clean no-primary control and similar results in adjacent sections (standard IHC practice). HPA reports medium staining in cerebellar Purkinje cells and caudate glial cells, while germinal-center cells in lymph node were not detected (HPA tissue IHC). LRRN1 is membrane-associated, and subcellular imaging places it mainly in the endoplasmic reticulum with additional Golgi and vesicle localisation (UniProt Q6UXK5 topology; HPA subcellular). Diffuse nuclear-only signal, section-edge concentration, necrotic debris, or staining reproduced without primary antibody warrants investigation (standard IHC practice). Check endogenous peroxidase and tissue pigment before calling weak DAB signal positive (standard IHC practice).
Boster reagents
Best LRRN1 / Leucine-rich repeat neuronal protein 1 IHC Antibodies
A12074-1 has real IHC data from paraffin-embedded rat brain (A12074-1 IHC image caption). Human, mouse, and rat are listed as reactive species (catalog reactivity); no IF image is supplied (catalog images).
Real IHC data
Anti-LRRN1 Antibody ®
Cat # A12074-1
A12074-1 is listed for IHC and for human, mouse, and rat reactivity (catalog applications and reactivity). Its figure shows chromogenic staining of paraffin-embedded rat brain after EDTA retrieval at pH 8.0 (A12074-1 IHC image caption).
Which to pick: For tissue IHC, choose A12074-1: its own paraffin-section image used 2 μg/ml primary antibody and DAB detection; the fixative is unreported (A12074-1 IHC image caption). No IF/ICC validation is supplied for A12074-1, so IF/ICC requires independent validation (catalog applications and images). For cross-species work, A12074-1 lists human, mouse, and rat reactivity, while its pictured IHC result covers rat only (catalog reactivity; A12074-1 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6UXK5 (LRRN1_HUMAN, Leucine-rich repeat neuronal protein 1).
- Human Protein Atlas. LRRN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LRRN1 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum. In addition localized to the Golgi apparatus, vesicles, centriolar satellites and basal body..
- Human Protein Atlas. LRRN1 antibody validation summary (1 antibodies).
- Knockdown of LRRN1 inhibits malignant phenotypes through the regulation of HIF-1α/Notch pathway in pancreatic ductal adenocarcinoma. Molecular therapy oncolytics 2021 — PMC8479295.
- Leucine-rich repeat neuronal protein-1 suppresses apoptosis of gastric cancer cells through regulation of Fas/FasL. Cancer science 2019 — PMC6609808.
- PubMed PMID:10819331 — UniProt-cited evidence.
- PubMed PMID:12880961 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LRRN1 (Leucine-rich repeat neuronal protein 1). Expected localisation General cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A12074-1). Validated IHC antibody: A12074-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Fallopian tube (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A12074-1); 2. Fixation: Image fixative and duration unreported (datasheet A12074-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A12074-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A12074-1); 8. Primary antibody: Rabbit anti-LRRN1, 2-5 μg/ml (datasheet A12074-1); 9. Primary incubation: Overnight at 4 °C (datasheet A12074-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA have medium consistency (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; epitope may be extracellular or cytoplasmic (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt Q6UXK5, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the endoplasmic reticulum. In addition localized…, Datasheet A12074-1, PMC8479295, PMC6609808. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LRRN1/immunohistochemistry-ihc-guide.